Yasumura S, Weidmann E, Hirabayashi H, Johnson J T, Herberman R B, Whiteside T L
Pittsburgh Cancer Institute, University of Pittsburgh School of Medicine, PA 15213.
Int J Cancer. 1994 May 1;57(3):297-305. doi: 10.1002/ijc.2910570302.
A human cytotoxic-T-lymphocyte (CTL) line capable of killing autologous tumor (AuTu) cell targets was established from peripheral-blood lymphocytes of a patient with squamous-cell carcinoma of the tongue. The cultured CTL were CD3+CD8+CD11b-HLA-DR+T cell receptor (TCR) alpha/beta+. When tested in 4-hr 51Cr-release assays against various lines of squamous-cell carcinoma of the head and neck (SCCHN) and a variety of non-squamous human tumor and normal cell targets, the CTL were found to lyse the autologous SCCHN cell line (PCI-50) and 7 allogeneic SCCHN lines: PCI-1, -2, -4A, -4B, -13, -30 and -38. Of these tumor cell lines, PCI-13, -30 and -38 shared HLA-A2 locus with the AuTu, PCI-50, while PCI-4A and -4B shared HLA-B44 with AuTu. Lysis of AuTu (A2+B44+), PCI-13 (A2+B44-) and PCI-4B (A2- B44+) by the CTL was efficiently inhibited by monoclonal antibodies (MAbs) to CD3, CD8, TCR alpha/beta or the major-histocompatibility-complex (MHC)-class-I antigens. MAbs to HLA-A2 antigens inhibited lysis of PCI-50 or PCI-13 targets by the CTL. In cold-target inhibition assays, unlabeled PCI-4B or PCI-13 cells inhibited CTL lysis of AuTu targets. The CTL incubated in the presence of the HLA-A2+ SCCHN PCI-50 or -13, but not an HLA-A2+ gastric carcinoma, produced TNF-alpha, IFN-gamma and GM-CSF. The CTL were tested for their TCR V beta gene expression by polymerase chain reaction (PCR). At week 10 in culture, the time of the highest AuTu cytotoxicity mediated by the CTL line, V beta 6 was expressed by 26% of T cells. Three clones, obtained by limiting dilution from 10-week CTL and selected for high cytotoxicity against AuTu, were found to be V beta6+. Further analysis of the specificity of these clones indicated lytic activity against PCI-13 (A2+B44-), but not PCI-4B (A2-B44+) targets. In 16-week cultures, which retained AuTu cytotoxicity as well as V beta 6 expression, TCR V beta 2 was also expressed at high frequency (29%), and AuTu-reactive clones were found to be V beta 2+. Our results indicate that at least 2 different CTL populations (V beta 6+ and V beta 2+) are able to recognize SCCHN-associated antigen(s) and that the V beta 6+ T cells are HLA-A2 restricted, while V beta 2+ T cells may be HLA-B44 restricted.
从一名舌鳞状细胞癌患者的外周血淋巴细胞中建立了一种能够杀伤自体肿瘤（AuTu）细胞靶标的人细胞毒性T淋巴细胞（CTL）系。培养的CTL为CD3 + CD8 + CD11b - HLA - DR + T细胞受体（TCR）α/β +。在针对各种头颈鳞状细胞癌（SCCHN）细胞系以及多种非鳞状人肿瘤和正常细胞靶标的4小时51Cr释放试验中，发现该CTL能够裂解自体SCCHN细胞系（PCI - 50）和7种同种异体SCCHN细胞系：PCI - 1、-2、-4A、-4B、-13、-30和-38。在这些肿瘤细胞系中，PCI - 13、-30和-38与AuTu、PCI - 50共享HLA - A2位点，而PCI - 4A和-4B与AuTu共享HLA - B44。针对CD3、CD8、TCRα/β或主要组织相容性复合体（MHC）I类抗原的单克隆抗体（MAb）可有效抑制CTL对AuTu（A2 + B44 +）、PCI - 13（A2 + B44 -）和PCI - 4B（A2 - B44 +）的裂解。针对HLA - A2抗原的MAb可抑制CTL对PCI - 50或PCI - 13靶标的裂解。在冷靶抑制试验中，未标记的PCI - 4B或PCI - 13细胞可抑制CTL对AuTu靶标的裂解。在HLA - A2 + SCCHN PCI - 50或-13存在下培养的CTL，但不是HLA - A2 +胃癌细胞，可产生肿瘤坏死因子-α（TNF - α）、干扰素-γ（IFN - γ）和粒细胞-巨噬细胞集落刺激因子（GM - CSF）。通过聚合酶链反应（PCR）检测CTL的TCR Vβ基因表达。在培养第10周时，即CTL系介导的对AuTu细胞毒性最高的时候，26%的T细胞表达Vβ6。通过对第10周的CTL进行有限稀释获得并选择对AuTu具有高细胞毒性的3个克隆，发现它们为Vβ6 +。对这些克隆的特异性进一步分析表明，它们对PCI - 13（A2 + B44 -）具有裂解活性，但对PCI - 4B（A2 - B44 +）靶标无裂解活性。在保留对AuTu细胞毒性以及Vβ6表达的16周培养物中，TCR Vβ2也高频表达（29%），并且发现对AuTu有反应的克隆为Vβ2 +。我们的结果表明，至少有两种不同的CTL群体（Vβ6 +和Vβ2 +）能够识别与SCCHN相关的抗原，并且Vβ6 + T细胞受HLA - A2限制，而Vβ2 + T细胞可能受HLA - B44限制。
